Culture medium and culture method for constructing liver tumor scaffoldless organoid

Abstract

The invention discloses a culture medium and a culture method for constructing a liver tumor scaffoldless organoid. The culture medium comprises the following components: Advanced DMEM / F12, FBS, diabody, N-2, Noggin, B-27, EGF, FGF-10, Y-27632, A83-01, SB202190, R-Spondin, N-acetylcysteine, Nicotinamide, Wnt3A, HGF and PrimocinTM. The culture method comprises the steps of adding primary tumor cells separated and obtained into the above-mentioned special culture medium for resuspension, inoculating into an ultra-low adsorption U-shaped well plate after counting, and centrifugally culturing until organoids are formed. The culture medium and the culture method can realize in-vitro proliferation of primary liver cancer cells, can quickly construct the liver tumor scaffoldless organoids, and can maintain long-term stable growth of the liver tumor organoids. The tumor organoids formed by the culture medium and the culture method retain heterogeneity of tumor tissues of patients, and are suitable for drug sensitivity detection, tumor generation and development related research of in-vitro liver cancer chemotherapeutic drugs.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a culture medium and a culture method for constructing liver tumor scaffold-free organoids. Background technique [0002] Hepatocellular carcinoma ranks second in the world's cancer mortality rankings, and it has the characteristics of high metastasis rate, low prognosis rate, and difficult treatment. Malignant tumor invasion and metastasis lead to tumor recurrence in patients, poor treatment effect and poor prognosis. Once a tumor cell is transformed into a malignant cell, its ability to invade and migrate becomes extremely strong. Tumor cells are not limited to the migration of single cells. In many tumor types, the commonly observed unit of invasion is a population of cells, the behavior of which determines malignant function. Two-dimensional (2-dimensional, 2D) cell culture has been widely used in biological research, but cells grown in two dimensions cannot accurately...

AI Technical Summary

Problems solved by technology

Two-dimensional (2D) cell culture has been widely used in biological research, but cells grown in two dimensions cannot accurately simulate the state in vivo

However, the current 3D culture technology of tumor organoids still has certain deficiencies (Science 364, 2019, 952–955.), such as: the shape of tumor models in different organs is not easy to distinguish, lack of representativeness; the culture period is long, and the culture price is relatively too high. High; organoid culture is mostly limited to matrix materials such as matrigel and hydrogel, which limit the gas exchange and substance metabolism between organoids and the outside world, and the exchange limitations seriously affect the absorption of nutrients required by organoids and removal of metabolic waste

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